| Part 1Objective: To explore the clinical significance of Wnt/β-catenin signaling pathway in rheumatoid arthritis(RA)patients,the expression levels of β-catenin,c-Myc and MMP-9 in peripheral blood and the changes of cytokines were observed.Methods : A total of 64 RA patients hospitalized in the Rheumatology Department of The Affiliated Hospital of Guilin Medical College from July2020 to July 2021 were selected.According to 28-joint disease activity score(DAS28),patients were divided into low disease activity group(n=17),moderate disease activity group(n=22)and severe disease activity group(n=25).At the same time,44 healthy subjects in the Health Examination Center were selected as healthy controls(HC).Age,gender,BMI,course of disease,morning stiffness time,tender joint count(TJC)and swollen joint count(SJC)were recorded in detail.ESR,CRP,RF,Anti-CCP,ALB and GLO were collected in the laboratory.The synovium of the joint of both hands was examined by musculoskeletal ultrasound.The protein expression levels of IL-1β,IL-6,IL-17 and TNF-α in peripheral blood serum were detected by ELISA.The m RNA expression levels of β-catenin,c-Myc and MMP-9 in peripheral blood were detected by Real-Time PCR.Results:(1)Compared with HC group,the levels of RF,Anti-CCP and GLO in RA group were significantly increased,while ALB was decreased(P<0.05).(2)Compared with HC group,the protein expression levels of IL-1β,IL-6,IL-17 and TNF-α in RA group were significantly increased(P<0.05).(3)Compared with HC group,m RNA expression levels of β-catenin,c-Myc and MMP-9 in peripheral blood of RA group were significantly increased(P<0.05).(4)The course of disease,morning stiffness time,TJC,SJC,ESR,CRP,RF,GLO and synovial greyscale ultrasound scores were positively correlated with disease activity in RA patients with different disease activity groups(P<0.05).There were no significant differences in Anti-CCP,ALB and synovial power doppler ultrasound scores among groups(P>0.05).(5)The protein expression levels of IL-1β,IL-6,IL-17,TNF-α and m RNA expression levels of β-catenin,c-Myc,MMP-9 in peripheral blood of RA patients with different disease activity groups were positively correlated with disease activity(P<0.05).(6)The m RNA expressions of β-catenin and MMP-9 in peripheral blood of RA patients were positively correlated with IL-1β,IL-6,IL-17 and TNF-α(P<0.05).The m RNA expression level of c-Myc was positively correlated with IL-1β,IL-6 and TNF-α(P<0.05).Conclusions: The expression levels of β-catenin,c-Myc and MMP-9 in peripheral blood of RA patients were significantly increased,suggesting that the Wnt/β-catenin pathway was abnormally activated and correlated with the disease activity of RA.Part 2Objective: Based on the experimental model of cultured collagen-induced arthritis(CIA)rat’s knee fibroblast-like synoviocytes(FLS),to investigate the effects of isoquercetin on proliferation of CIA-FLS and Wnt/β-catenin signaling pathway.Methods :(1)CIA-FLS was identified by immunohistochemistry.(2)CCK-8 detected the proliferation of CIA-FLS in IQC groups with different concentration gradients,Wnt inhibitory factor-1(Wif-1)positive control group and Li CL negative control group.(3)The levels of TNF-α and IL-6 in the supernatant of CIA-FLS treated with different concentrations of IQC,Wif-1 and Li CL were detected by ELISA.(4)The m RNA expressions of IL-6,TNF-α,c-Myc,β-catenin and MMP-9 in cells treated with different concentrations of IQC,Wif-1 and Li CL were detected by Real-Time PCR.(5)Western Blot was used to detect the protein expressions of c-Myc,β-catenin and MMP-9 in cells treated with IQC,Wif-1 and Li CL at different concentrations.Results:(1)IQC inhibited CIA-FLS proliferation and antagonized Li CL proliferation.(2)IQC can reduce the protein and m RNA expression levels of IL-6 and TNF-α in CIA-FLS.(3)IQC can reduce the protein and m RNA expression levels of β-catenin,c-Myc and MMP-9 in CIA-FLS.Conclusions: Isoquercitrin may inhibit the proliferation of CIA-FLS and the release of inflammatory factors through Wnt/β-catenin signaling pathway. |
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